Light-stimulated release of cargo from oligonucleotides

Abstract

The invention provides oligonucleotide conjugates including a photolabile crosslinker attached to a cargo moiety, e.g., a therapeutic or diagnostic agent. The invention further provides reagents useful in the preparation of such conjugates and methods of their use.

What is claimed is: 
     
       1. An oligonucleotide conjugate of the formula: 
       
         
           
           
               
               
           
         
       
       wherein Oligo is an oligonucleotide of 2-1000 nucleotides in length, Ai is the residue of a conjugation reaction, A 2  is an amine reactive leaving group or —NHX, L is an optional linker, Y is H or C1-10 alkyl, and X is a cargo moiety. 
     
     
       2. The conjugate of  claim 1 , wherein L is present and amido, C1-10 alkylene, or C1-20 polyalkeneoxide. 
     
     
       3. The conjugate of  claim 1 , wherein L is present and C2-C20 polyethylene glycol. 
     
     
       4. The conjugate of  claim 1 , wherein Ai is triazolyl, disulfide, cyclohexenyl, amido, thioamido, acetal, ketal, or sulfonamido. 
     
     
       5. The conjugate of  claim 1 , wherein Y is C1-10 alkyl. 
     
     
       6. The conjugate of  claim 1 , wherein Y is methyl. 
     
     
       7. The conjugate of  claim 1 , wherein A 2  is the amine reactive group. 
     
     
       8. The conjugate of  claim 7 , wherein the amine reactive group is p-nitrophenoxyl, N-hydroxysuccinimidyl, halide, pentafluorophenoxyl, or imidazolyl. 
     
     
       9. The conjugate of  claim 1 , wherein A 2  is NHX. 
     
     
       10. The conjugate of  claim 9 , wherein X is a therapeutic or diagnostic agent. 
     
     
       11. The conjugate of  claim 1 , having the formula: 
       
         
           
           
               
               
           
         
       
       wherein Oligo is an oligonucleotide of 2-100 nucleotides in length, T is a triazolyl linker formed from the reaction of an azide with an alkyne, X is a cargo moiety, and n is an integer from 1-10. 
     
     
       12. A crosslinker of the formula: 
       
         
           
           
               
               
           
         
       
       wherein A 3  is a conjugating moiety, A 4  is an amine reactive leaving group, L is C2-C20 polyethylene glycol, and Y is H or C1-10 alkyl. 
     
     
       13. The crosslinker of  claim 12 , wherein A 3  is azido, alkynyl, alkenyl, thiol, halide, boronic acid, hydroxyl, carboxyl, formyl, or ketone. 
     
     
       14. A crosslinker of the formula: 
       
         
           
           
               
               
           
         
       
       wherein A 3  is a conjugating moiety, A 4  is an amine reactive leaving group, L is an optional linker, and Y is C1-10 alkyl. 
     
     
       15. The crosslinker of  claim 14 , wherein Y is methyl. 
     
     
       16. The crosslinker of  claim 12 , wherein A 4  is p-nitrophenoxyl, N-hydroxysuccinimidyl, halide, pentafluorophenol, or imidazolyl. 
     
     
       17. The crosslinker of  claim 12 , having the formula: 
       
         
           
           
               
               
           
         
       
       where n is an integer from 1-10. 
     
     
       18. A DNA construct comprising a three-dimensional DNA cage structure for housing a cargo moiety, wherein the cargo moiety is attached to the cage structure via the oligonucleotide conjugate of  claim 1 . 
     
     
       19. A method of delivering a cargo moiety, the method comprising providing a conjugate of  claim 1  and irradiating the conjugate with light to release the cargo moiety. 
     
     
       20. The method of  claim 19 , wherein the conjugate is internalized within a cell prior to irradiation. 
     
     
       21. The method of  claim 19 , wherein the cargo is a therapeutic or diagnostic agent. 
     
     
       22. A pharmaceutical composition comprising a conjugate of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
       23. The crosslinker of  claim 14 , wherein A 3  is azido, alkynyl, alkenyl, thiol, halide, boronic acid, hydroxyl, carboxyl, formyl, or ketone. 
     
     
       24. The crosslinker of  claim 14 , wherein A 4  is p-nitrophenoxyl, N-hydroxysuccinimidyl, halide, pentafluorophenol, or imidazolyl.